CN105031636B - A kind of Aeromonas hydrophila and Aeromonas veronii bivalent inactivated vaccine and preparation method - Google Patents
A kind of Aeromonas hydrophila and Aeromonas veronii bivalent inactivated vaccine and preparation method Download PDFInfo
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- CN105031636B CN105031636B CN201410468192.9A CN201410468192A CN105031636B CN 105031636 B CN105031636 B CN 105031636B CN 201410468192 A CN201410468192 A CN 201410468192A CN 105031636 B CN105031636 B CN 105031636B
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Abstract
The present invention provides a kind of Aeromonas hydrophila and Aeromonas veronii bigeminy oral slow-releasing microsphere vaccine and preparation methods, belong to field of biological pharmacy.Using sodium alginate as wall material, Aeromonas hydrophila and Aeromonas veronii inactivate full bacterium as core material, and Aeromonas hydrophila and Aeromonas veronii bigeminy oral slow-releasing microsphere vaccine are prepared using emulsion process.Pond crucian carp is immunized using the Aeromonas hydrophila and Aeromonas veronii-sodium alginate microcapsule oral vaccine, the activity of serum enzyme that can be significantly improved, leukocytes phagocytic is active, and the relative protection ratio to pond crucian carp is：46.7%.Micro-capsule is stablized in the case where simulating gastrointestinal conditions, shows preferable burst release and slow release；Bacterial antigen keeps good in micro-capsule, and vaccine safety is preferable with storage stability.
Description
Technical field
The present invention relates to animal vaccine technologies of preparing, and in particular to a kind of Aeromonas hydrophila and Aeromonas veronii bigeminy
Inactivated vaccine and preparation method.The vaccine of the present invention is primarily adapted for use in freshwater fish.
Background technology
Aeromonas hydrophila and Aeromonas veronii (Aeromonas hydrophil) belong to vibrionaceae, Aeromonas,
It is widely present in fresh water, soil, sewage, mud and excrement, there is pathogenic bacterial strains and non-pathogenic bacterial strains.Pathogenic bacterium
Strain pattern of infection is very extensive, the outburst of soft-shelled turtle, shrimp, crab, the frog, duck etc. can be caused to infect, this bacterium of human infection can cause food poisoning
It is infected with soft tissue injury, is the main pathogenic fungi of freshwater fish.It is that China breeds fish history by the microbial bacterial septicemia
On endanger fish most species, the waters classification that endangers that fish the range of age is maximum, Prevalent district is most wide, harm is breeded fish at most, cause to damage
Lose a kind of one of acute infectious disease of most serious.
At present to the medicine of this disease, predominantly antibiotic and chemicals, a large amount of be used for a long time is also easy to produce persister,
And chemicals can cause to remain, and pollute water quality, destroy the ecological balance.So it is prevention and control to produce effective vaccine
The key of this disease.Evaluation of Aeromon As Hydrophila Vaccine and its relevant technologies achieve one in the research and extension application of recent decades
The gratifying achievement of series.The type of vaccine also tends to diversification, appearance and hair along with molecular biology and genetic engineering
, there is the novel fish vaccine such as such as nucleic acid vaccine, recombinant vaccine, synthetic peptide vaccine in exhibition.But these vaccines there is also
Some problems, as immune effect is unstable, safety is poor, technology of preparing is more difficult, cost is higher, it is rationally effectively convenient to lack
Administration way etc., therefore, also there are certain difficulties for the extensive use of these vaccines in actual production.
Aeromonas hydrophila and Aeromonas veronii be it is heterologous, due to Physiological-biochemical Characters between different strains, serotype,
Genotype be it is discrepant, it is pathogenic also due to it expression virulence factor on difference and it is different, thus, in the general of vaccine
Limited on and.
Invention content
In order to solve the deficiencies in the prior art, the present invention provides a kind of Aeromonas hydrophila and Aeromonas veronii bigeminy is gone out
Live vaccine and preparation method.
The technical scheme is that：Applicant is prepared for a kind of Aeromonas hydrophila and the inactivation of Aeromonas veronii bigeminy
Vaccine, the inactivated vaccine contain preserving number for CCTCC NO:The antigen and preserving number of the Aeromonas hydrophila of M 2013566 is
CCTCC NO:The antigen of the Aeromonas veronii of M 2013565；The antigen has inactivated.The antigen of the Aeromonas hydrophila
Antigen volume ratio with Aeromonas veronii is 1:1.
Applicant additionally provides the preparation method of a kind of Aeromonas hydrophila and Aeromonas veronii bivalent inactivated vaccine, packet
Include following steps：It is CCTCC NO by preserving number:The Aeromonas hydrophila of M 2013566 and preserving number are CCTCC NO:
After the Aeromonas veronii Multiplying culture of M2013565, collect thalline, add in phosphate buffer (PBS) it is dense to bacterium be 1 ×
108Cfu/mL, then adds in final concentration of 0.2% formalin, 28 DEG C of shaking table cultures for 24 hours, Aeromonas hydrophila and Vickers
The content of Aeromonas is 50%, and the inactivated vaccine of bigeminy bacterium is made.
The isolation and identification method of above-mentioned antigen bacterial strain is as described below：
Aeromonas hydrophila and Aeromonas veronii bacterial strain, applicant is from the sick fish body that bacteremic septicemia is suffered from by Xinxiang City
Separation, wherein 32 plants of Aeromonas hydrophila, 20 plants of Vickers gas unit cell power, freeze-drying preserve.According in challenge test, every group of crucian carp
The death condition of fish, and the characteristics of combine a Strain Virulence factor, has filtered out that virulence factor is more, and the bacterial strain of strong toxicity is as epidemic disease
Seedling prepares spare bacterial strain:Aeromonas hydrophila XDMG (1) Aeromonashydrophil XDMG (1), LD50For 1.5 ×
105cfu/mL.Aeromonas veronii XDLG (1) Aeromonasveronii XDLG (1), LD50It is 4.5 × 105cfu/mL。
More than bacterial strain was preserved in China typical culture collection center (CCTCC), preservation address on November 11st, 2013：Chinese lake
Wuhan University of Bei Sheng Wuhan City.Its preserving number is respectively CCTCC NO:M 2013566 and CCTCC NO:M 2013565.Sampling
Time be in July, 2010, place:Xinxiang, Weihui City East Lake are taken respectively from the liver in pseudorasbora parva and silver carp.
The present invention chooses the Aeromonas hydrophila XDMG (1) of more north Henan virulence factor, immunogenicity height, strong toxicity
Aeromonashydrophil XDMG (1) and Aeromonas veronii XDLG (1) Aeromonasveronii XDLG (1) are made two
Join inactivated vaccine.Vaccine is prepared using formalin-inactivated, crucian (30~35g) is injected intraperitoneally, carries out serum antibody titer
Detection, pathological section analysis and protest test.The result shows that crucian detects aggegation on the 3rd week after injecting immune
Antibody peaked in the 6th week agglutination titer, and control group does not detect antibody in whole experiment process；Pathological section
It also indicates that, which can generate good protective effect to crucian target organ；In protest test, the immune guarantor of immune group
Shield rate is up to 100%, and immune period is up to 6 months or more.Aeromonas hydrophila XDMG (1) Aeromonashydrophil
XDMG (1) and Aeromonas veronii XDLG (1) Aeromonasveronii XDLG (1) inactivated vaccines, which have crucian, significantly exempts from
Epidemic disease protective effect, can be as the vaccine of prevention bacterial septicemia infection.
Description of the drawings
Fig. 1 is aer (1), alt (2), ahp (3) and β-hly in 3 plants of A.Aeromonashydrophil XDMG (1)
(4) the PCR amplification result of gene.
2 source of fish bacterial strain of the typical sick fish isolated strains of tri- kinds of Fig. 2, four kinds of source seedling strains virulence gene relative expression 1,3 water source
Bacterial strain.
Fig. 3 each group serum antibody titer average values.
The histotomy of the liver of experimental group and control group fish, spleen, kidney, intestinal tissue under Fig. 4 light microscopics.
Specific embodiment
In order to make the present invention it is more readily appreciated that elaborating with reference to specific embodiment to the present invention.This single implementation
Example is not limitation of the present invention.
Embodiment
1 material
1.1 bacterial strain
Aeromonas hydrophila and Aeromonas veronii bacterial strain are detached in the sick fish body for suffering from bacteremic septicemia from Xinxiang City,
Wherein 32 plants of Aeromonas hydrophila, 20 plants of Vickers gas unit cell power, freeze-drying preserve.
1.2 experimental animal
Healthy crucian is purchased from Xinxiang suburbs fishing ground, and every fish quality is about 30g, and length is 12~15cm.Use preceding elder generation
It raises and train 1 week, inflates in laboratory, control 28 DEG C or so of water temperature, periodically remove contamination and change water, feeding.
1.3 culture medium
The preparation of LB nutrient broths:Beef extract 3.0g, peptone 10.0g, sodium chloride 5.0g, potassium dihydrogen phosphate 1.0g steam
Distilled water 1000mL, mixing dissolve by heating, and pH value is adjusted to be dispensed, 112kPa high pressure sterilizations 15min to 7.6.
The preparation of LB agar plates:Beef extract 3.0g, peptone 10.0g, sodium chloride 5.0g, potassium dihydrogen phosphate 1.0g, fine jade
Fat 15g, distilled water 1000mL, mixing dissolve by heating, and adjusting pH value, packing, 112kPa high pressure sterilization 15min are cooled to 7.6
45 DEG C of sterile pour plates.
2 methods
The screening of 2.1 vaccine strains
2.1.1 virulence factor PCR detections and parting
4 kinds of primers：Gas lysin (aer), beta hemolysin (β-hly), cell excitement enterotoxin (alt), the extracellular egg of serine
White enzyme (ahp) is synthesized by Shanghai Sheng Gong biotech companies, and sequence is shown in Table 1.PCR reactions, mould are carried out using 25 μ L systems
A concentration of 30ng/ μ L of plate, Ago-Gel carry out electrophoresis, detect 4 kinds of virulence factors.
14 kinds of virulence gene P C R amplimers sequences of table and target fragment size
2.1.2 the real-time quantitative analysis of virulence factor
The real-time quantitative primer (being shown in Table 2) of 4 kinds of virulence genes of design.The bacterial strain more than virulence factor is filtered out, extracts RNA
When row reverse transcription synthesis cDNA, reacted on PR0961101428 real-time quantitative PCR instruments using 10 μ L systems, detection 4
The relative expression quantity of kind virulence factor.
The real-time primer sequence of 24 kinds of virulence genes of table and target fragment size
2.1.3 half lethal concentration (LD50) measure
After the bacterial strain activation that laboratory preserves, it is inoculated in 28 DEG C of shaking table culture 16h of nutrient broth.Culture utilizes blood bead
Tally counts, 5 groups of the experiment component of every plant of bacterium, every group of 10 tail fishes, by bacterium solution with 10-1, 10-2, 10-3, 10-4Doubling dilution, often
Crucian is injected intraperitoneally in dilution bacterium solution, and 0.2mL is penetrated, while set saline control per endnote.Experiment fish is supported sterilized in
In aquarium, tap water has been aerated for 24 hours, and inflation controls 28 DEG C or so of water temperature, periodically removes contamination and change water, feed food.Record daily
The death condition of every group of fish calculates half lethal concentration (LD with reference to Reed-Muench methods50)。
The preparation of 2.2 full bacterium inactivated vaccines
2.2.1 strain source
XDLG(1)；XDMG(1).
2.2.2 actication of culture is cultivated with expanding
The aseptically a small amount of lawn of picking from Aeromonas hydrophila and Aeromonas veronii opportunistic pathogen kind dry powder, inoculation
In nutrient broth test tube after shaking table culture, setting-out is inoculated in agar plate, and 28 DEG C of culture 18-24h grow bacterium colony, through inspection
After looking into no miscellaneous bacteria, then picking single bacterium colony is inoculated in and contains in 200mL nutrient broth medium triangular flasks (pH 7.2), 28 DEG C of oscillation trainings
(frequency of oscillation is 120 beats/min -130 beats/min) is supported, culture 18-24h is for use.
Strain can continue to be enlarged culture with triangular flask, inoculum concentration 10%, cultural method with strain culture, then
Add in final concentration of 0.2% formalin, 28 DEG C of shaking table cultures for 24 hours, the content of Aeromonas hydrophila and Aeromonas veronii
It is 50%, the inactivated vaccine of bigeminy bacterium is made.But during with shaking table culture, yield is relatively low, therefore generally uses fermentation tank culture.
Seed tank culture
Fermentation tank sterilizing postcooling can be inoculated with to 28 DEG C.The expansion bacterium that will aseptically be made in 1% ratio
Liquid is inoculated in be cooled in 28 DEG C of seeding tank and be cultivated.28 DEG C of temperature control after inoculation, pressure are 0.05MPa ventilatory capacities 1：1 He
18-24h is cultivated under 220r/min stirring conditions.22h starts to sample one-time detection every 2h after inoculation, when thalli growth is neat；
In exponential phase；Bacteria containing amount is 1 × 108More than cfu/mL；Without living contaminants；pH 7.0-7.5.It can culture transferring extremely fermentation
Fermented and cultured in tank.
Fermentation tank culture
Aseptically the inoculum of seeding tank by 1% inoculative proportion is inoculated in and is cooled to 28 DEG C of fermentation
It is cultivated in tank, condition of culture and Testing index are same as above.When bacteria containing amount reach production needs after, you can with fermentation tank bacterium solution into
Row inactivation.
2.2.3 inactivation
Inoculum in fermentation tank is added in into formalin, final concentration of 0.2%, it inactivates for 24 hours under the conditions of 28 DEG C, is going out
It is sufficiently stirred 2-3 times during living.Bacterium solution after inactivation is Fish Aeromonas Hydrophila and Aeromonas veronii inactivation bigeminy
Vaccine.
2.3 vaccine Sterility testings and safety testing
The vaccine spread plate of above-mentioned preparation, 28 DEG C of culture 48h are taken, observation whether there is bacterium colony appearance.
It takes the vaccine of above-mentioned preparation that 30 tail of crucian is injected intraperitoneally, 0.2mL is penetrated per endnote, the reaction of observation experiment crucian and dead
Die situation.
Inactivation, safety and effect are detected into qualified vaccine aseptically, 50mL is opened in packing or 100mL is sterilized
In the glass or plastic bottle of processing, product is packed in labelling, and 4 DEG C stored refrigerated, and the term of validity is 90 days.
The immunity inoculation of 2.4 crucians
Experimental group and each 80 tail crucian of control group, experimental group are injected intraperitoneally full bacterium inactivation solution, 0.2mL are penetrated per endnote；It is right
According to a group physiological saline for intraperitoneal injection equivalent.Inflation daily, feeds food, and water temperature is controlled at 28 DEG C or so.Record the dead of each group
Die situation.
2.5 Serum Antibody Detection
After crucian is immunized, take 5 tails at random from each group fish weekly, syringe arteria caudalis/venous blood sampling, blood collection in from
Heart pipe makes natural subsidence at its 4 DEG C, detaches serum.After 96 orifice plates add in 50 μ L of physiological saline per hole, it is dilute to be separately added into 2 multiple proportions
Release immune fish serum and and control fish serum, then add in the bigeminy bacterium solution of inactivation as antigen, culture plate is in 28 DEG C of insulating boxs
1h judges result after placing into 4 DEG C of refrigerator overnights.
2.6 pathological sections are analyzed
After crucian is immunized 21 days, the fish of random picking experimental group and control group, with 50LD50Aeromonas hydrophila and Vickers gas
Monad is attacked, and solution takes its liver, spleen, kidney and intestinal tissue and fixes after 12h, is embedded, is sliced through freezing microtome,
HE is dyed, and dimethylbenzene is transparent, and after neutral gum is sealed up for safekeeping, in optical microphotograph Microscopic observation, simultaneously contrast experiment's group is each with fish in control group
The variation of histoorgan.
2.7 protest test
After first immunisation 21 days, every group of random 30 tail fish of picking, with 50LD50Aeromonas hydrophila and Aeromonas veronii
Attack equally takes the mode of intraperitoneal injection, cultivates 10d under similary environment, each group death toll is observed and recorded daily, according to public affairs
Formula calculates the immune protective rate of fish：
Immunoprotection (%)=(the control group fingerling death rate-test group fingerling death rate)/control group fingerling death rate ×
100%
The measure of 2.8 immune periods
After first immunisation 6 months, every group of random 30 tail fish of picking, still with 50LD50Aeromonas hydrophila and Vickers unit cell
Bacterium is attacked, and cultivates 10d under similary environment, observes and records each group death toll daily, and the immune protective rate of fish is calculated according to formula.
3 results
The screening test of 3.1 vaccine strains
3.1.1 virulence factor PCR detections and parting
There was only 3 kinds of virulence genotypes in 52 plants of bacterium：aer+alt+ahp+hly+、aer+alt-ahp+hly+、aer+alt+ahp-
hly+-.The aer that wherein 4 kinds of virulence factors have+alt+ahp+hly+Type accounts for 67.3% (35/52).Fig. 1 is 3 kinds of virulence genotypes
Representative strain PCR amplification the result is shown in Figure 1.
3.1.2 the real-time quantitative analysis of virulence factor
Utilize the aer containing 4 kinds of virulence genotypes+alt+ahp+hly+Representative strains carry out real-time quantitative to 4 kinds of virulence genes
Analysis.Using 16SRNA as internal reference, using typical sick fish separation strains as control, quantitative analysis is carried out to the source of fish, water source bacterial strain respectively,
The relative expression quantity of representative strain is shown in Fig. 2.Typical disease 4 kinds of virulence gene relative expression quantity highests of fish separation strains and significant difference.
3.1.3 half lethal concentration (LD50) measure
The bacterial strain activation that 4 kinds of virulence factor relative expression quantities are high is selected, has carried out half lethal concentration LD50Measure.Root
According in challenge test, the death condition of every group of crucian, and the characteristics of combine a Strain Virulence factor, filtered out virulence factor
More, the bacterial strain of strong toxicity prepares spare bacterial strain as vaccine:Aeromonas hydrophila, LD50It is 1.5 × 105cfu/mL.Vickers gas
Monad, LD50It is 4.5 × 105cfu/mL.More than bacterial strain was preserved in Chinese Typical Representative culture guarantor on November 11st, 2013
Tibetan center (CCTCC), preserving number are respectively CCTCC NO:M 2013566 and CCTCC NO:M 2013565.
3.2 vaccine Sterility testings and safety testing
It is sterile to drop out now, it was demonstrated that vaccine inactivation is complete after inactivated vaccine spread plate culture 48h is used in experiment.
In animal safety experiment, it is no different paradoxical reaction after vaccine injection crucian, it was demonstrated that vaccine is safety non-toxic.
3.3 Serum Antibody Detection
Every fish serum agglutinating antibody potency and its average value are shown in Fig. 3.From the figure 3, it may be seen that crucian is after injecting immune,
It detects agglutinating antibody within 3 weeks, peaks in the 6th week agglutination titer, and control group does not detect in whole experiment process
To antibody.
3.4 pathological sections are analyzed
Experimental group is shown in Fig. 4 with control group pathological section, as shown in figure 4, figure A is in for the visible liver cell of immune group primary culture
Cerioid, it is that the visible spleen essence red pulp of immune group fish spleen and white pulp are spaced to arrange uniformly regular (shown in arrow) figure B,
Without apparent boundary, (the arrow institute such as red blood cell, granulocyte, lymphocyte of different developmental phases is filled in desmachyme
Show).Figure C is immune group fish kidney, it is seen that its form normal in size, skin medullary substance distinct, for sinus renalis without separation, renal tubule is normal
(shown in arrow)；Figure D be immune group fish intestinal tissue, Epithelial cell, lymphocyte, mucosal inner layer, submucosa cell
Form is normal (shown in arrow)；Scheme the liver that E is control group fish, liver cell packet slurry is loose and apparent ballooning degeneration occurs, after
There is a large amount of necrosis of liver cells in phase and lymphocytic infiltration, red blood cell enlargement, rupture, dissolving, capillary, thin vessels are broken
Damage, tube wall is flat, endotheliocytic swelling, denaturation, significant hemolysis, it is seen that brown color hematogenous pigment is deposited (shown in arrow)；Scheme F
Spleen for control group fish, it is seen that spleen red pulp is difficult to recognize with white pulp, and minibody structure is unclear, and histocyte largely dissolves, is bad
Extremely, hematogenous pigment deposition is the most notable (shown in arrow)；Scheme the kidney that G is control group fish, it is seen that Malpighian corpuscle necrosis, glomerulus
Blister cavities becomes larger, renal cells swelling, denaturation, necrosis, disintegration (shown in arrow)；Scheme the enteron aisle that H is control group fish, it is seen that
Intestinal villus necrosis comes off, and the necrosis of intestinal villus epithelial cell comes off, a large amount of hyperplasia of goblet cell (shown in arrow).Illustrate vaccine pair
Crucian produces protective effect, and when in Aeromonas hydrophila and Aeromonas veronii intrusion fish body, the immune system of fish can be known
Not and pathogen is eliminated, in case bacterium causes to damage to tissue.
3.5 protest test
Inactivated vaccine shows the protest test (being shown in Table 3) of crucian the vaccine can generate crucian protection well
Effect, immune protective rate have reached 100%；Non-immune control group crucian is all dead, is recovered to and attacks from dead fish internal organ
Strain.
3 experimental group of table and the death condition of control group protest test fish
The measure of 3.6 immune periods
Immunoprotection result (being shown in Table 4) display after 6 months, which is still up to 97%, and is not immunized
Control group crucian it is all dead, be recovered to from dead fish internal organ and attack strain, illustrate that the vaccine is long to the immune period of crucian
Up to 6 months or more.
Experimental group and the death condition of control group protest test fish after table 46 months
This research is suffered from naturally from Xinxiang area to be detached in the case of bacterial septicemia, filters out velogen strain, using formaldehyde
The method of inactivation prepares Aeromonas hydrophila and Aeromonas veronii inactivated vaccine, and crucian is immunized with this vaccine, due to being
The whole-bacterial-vaccine of inactivation, it is not only safe and non-toxic, but also most antigenic components of bacterium are saved, and using virulence factor most
More, the most strong bacterial strain of toxicity is as vaccine strains, so immanoprotection action can be generated to more kinds of velogen strains.Serum antibody is examined
Explanation is tested in test, in immune response, produces antibody, specific immunity has played important function.It can from pathological section
Go out, Aeromonas hydrophila can invade the tissues such as liver, spleen, kidney, the enteron aisle of fish, and cause serious damage, and vaccine can
To be killed before bacterium enters these tissues, the safety of fish body is protected well.
The basic principles, main features and the advantages of the invention have been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent thereof.
Claims (2)
1. a kind of Aeromonas hydrophila and Aeromonas veronii bivalent inactivated vaccine, which is characterized in that the inactivated vaccine contains guarantor
Tibetan number is CCTCC NO:The antigen and preserving number of the Aeromonas hydrophila of M 2013566 are CCTCC NO:The dimension of M 2013565
The antigen of family name Aeromonas；The antigen has inactivated.
2. a kind of Aeromonas hydrophila according to claim 1 and Aeromonas veronii bivalent inactivated vaccine, feature exist
In the antigen of the Aeromonas hydrophila and the antigen volume ratio of Aeromonas veronii are 1:1.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1931364A (en) * | 2006-09-08 | 2007-03-21 | 陈刚 | Prepn process of oral vaccine for antagonizing bacterial diseases of aquatic animal |
| CN101862308A (en) * | 2010-03-16 | 2010-10-20 | 南京农业大学 | Aeromonas hydrophila micro-capsular oral vaccine |
| CN102139102A (en) * | 2011-04-01 | 2011-08-03 | 上海海洋大学 | Preparation method of microcapsule vaccines capable of resisting infection of aeromonas hydrophila |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1931364A (en) * | 2006-09-08 | 2007-03-21 | 陈刚 | Prepn process of oral vaccine for antagonizing bacterial diseases of aquatic animal |
| CN101862308A (en) * | 2010-03-16 | 2010-10-20 | 南京农业大学 | Aeromonas hydrophila micro-capsular oral vaccine |
| CN102139102A (en) * | 2011-04-01 | 2011-08-03 | 上海海洋大学 | Preparation method of microcapsule vaccines capable of resisting infection of aeromonas hydrophila |
Non-Patent Citations (1)
| Title |
|---|
| 银鲫口服嗜水气单胞菌疫苗的免疫和免疫组化研究;李新华等;《水生生物学报》;20070120;第31卷(第01期);125-130 * |
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